An HpaII/MspI-PCR assay to measure methylation of DNA in Hoplosternum littorale (Callichthyidae, Siluriformes) from a polluted environment in the central Amazon basin

Silva, Francijara Araújo da; Feldberg, Eliana; Goll, Leonardo Gusso; Carvalho‐Zilse, Gislene Almeida; Gross, Maria Cláudia; Schneider, Carlos Henrique; Matoso, Daniele Aparecida

doi:10.1016/j.eti.2019.100354

Cited by 2 publications

(2 citation statements)

References 27 publications

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“…Moreover, we used the colorectal cancer cell line HT29 with hypermethylated genomic DNA [ 24 ]. The amount of demethylated DNA was detected using methyl-sensitive restriction (MSRE) assay based on the intensity of the HpaII-digested genome DNA cleavage products, as described in Da Silva et al [ 25 ]. We found a decrease in the intensity of HpaII-digested product by 33% after the treatment of Ca Ski cells with CBL0137 (0.5 μM), whereas there were no significant differences between experimental and control samples in HT29 cells after CBL0137 treatment (0.05 μM) ( Figure 1 C).…”

Section: Resultsmentioning

confidence: 99%

Insights into the Mechanism of Curaxin CBL0137 Epigenetic Activity: The Induction of DNA Demethylation and the Suppression of BET Family Proteins

Maksimova,

Popova,

Prus

et al. 2023

IJMS

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The development of malignant tumors is caused by a complex combination of genetic mutations and epigenetic alterations, the latter of which are induced by either external environmental factors or signaling disruption following genetic mutations. Some types of cancer demonstrate a significant increase in epigenetic enzymes, and targeting these epigenetic alterations represents a compelling strategy to reverse cell transcriptome to the normal state, improving chemotherapy response. Curaxin CBL0137 is a new potent anticancer drug that has been shown to activate epigenetically silenced genes. However, its detailed effects on the enzymes of the epigenetic system of transcription regulation have not been studied. Here, we report that CBL0137 inhibits the expression of DNA methyltransferase DNMT3a in HeLa TI cells, both at the level of mRNA and protein, and it decreases the level of integral DNA methylation in Ca Ski cells. For the first time, it is shown that CBL0137 decreases the level of BET family proteins, BRD2, BRD3, and BRD4, the key participants in transcription elongation, followed by the corresponding gene expression enhancement. Furthermore, we demonstrate that CBL0137 does not affect the mechanisms of histone acetylation and methylation. The ability of CBL0137 to suppress DNMT3A and BET family proteins should be taken into consideration when combined chemotherapy is applied. Our data demonstrate the potential of CBL0137 to be used in the therapy of tumors with corresponding aberrant epigenetic profiles.

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Section: Resultsmentioning

confidence: 99%

Insights into the Mechanism of Curaxin CBL0137 Epigenetic Activity: The Induction of DNA Demethylation and the Suppression of BET Family Proteins

Maksimova,

Popova,

Prus

et al. 2023

IJMS

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“…These isoschizomers are used in pairs for instance HpaII/MspI and recognize the same 5′-CCGG site. The main differences between them is sensitivity to methylation of cytosine [44]. MspI is not sensitive to methylation and remains undigested fragment which cannot be amplified in the PCR process.…”

Section: Hpaii/mspi-pcrmentioning

confidence: 99%

DNA Methylation Tools and Strategies: Methods in a Review

Niya

Roshanzamir

Mortazavi³

2019

Asian Pac J Cancer Biol

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DNA methylation is known as an important epigenetic change in plants and vertebrates genome. In this process, the methyl group transferred by DNA methyl transferase enzymes to cytosine at carbon residue 5 often in the CpG dinucleotide context. DNA methylation plays an important role in the natural development of the organism, genome stability maintenance and processes such as genomic imprinting and chromosome X inactivation in mammals. In addition, changes in DNA methylation pattern have seen in many diseases, including cancer. Analysis of DNA methylation has been useful for rapid disease diagnosis and progression. In recent decades, a revolution has taken place in the methods of DNA methylation analysis, and it is possible to study the pattern of gene methylation at a widespread, short and high resolution level. These methods can be divided into three general categories: (1) cut-based methods by methylation-sensitive enzymes; (2) sodium bisulfide based methods; (3) antibody based methods. Since the existence of different methods makes it difficult to select the appropriate approach, in this review, a number of common methods for examining the methylation pattern with the advantages and disadvantages will be discussed.

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An HpaII/MspI-PCR assay to measure methylation of DNA in Hoplosternum littorale (Callichthyidae, Siluriformes) from a polluted environment in the central Amazon basin

Cited by 2 publications

References 27 publications

Insights into the Mechanism of Curaxin CBL0137 Epigenetic Activity: The Induction of DNA Demethylation and the Suppression of BET Family Proteins

Insights into the Mechanism of Curaxin CBL0137 Epigenetic Activity: The Induction of DNA Demethylation and the Suppression of BET Family Proteins

DNA Methylation Tools and Strategies: Methods in a Review

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